Method and System for Liquid Chromatograph with Compressibility and Viscosity Monitoring to Identify Fluids

ABSTRACT

A system for providing a solvent or reagent to a liquid chromatography system comprises: a valve comprising a common port and a plurality of other ports, configurable such that the common port may be fluidically coupled to any one of the other ports; a pump fluidically coupled to the common port of the valve; a plug configured to block flow through a first one of said other ports of the valve; a container containing the solvent or reagent, said container fluidically coupled to a second one of said other ports of the valve; a fluid tubing line having a known resistance to fluid flow fluidically coupled to a third one of said other ports of the valve; and a pressure gauge configured to measure fluid pressure within the pump, wherein the solvent or reagent is provided to the liquid chromatography system by a fourth one of the other ports.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is the U.S. National Stage Application, under 35 U.S.C. 371, of International Application PCT/US2011/058230 having an international filing date of Oct. 28, 2011, which claims the benefit of the filing date, under 35 U.S.C. 119(e), of U.S. Provisional Application 61/408,046, filed on Oct. 29, 2010.

TECHNICAL FIELD

This invention generally relates to chromatography, and more particularly to an automated apparatus and method for monitoring the correctness of the installation and placement of solvents, mobile phases or other reagents within a chromatograph instrument or system.

BACKGROUND ART

Liquid chromatography (LC) is well-known in the fields of chemical separation, compound purification and chemical analysis. A liquid chromatograph generally includes a separation column that comprises a capillary tube that is packed with a permeable solid material that either is, itself, a chromatographic stationary phase or otherwise comprises or supports a chromatographic stationary phase. A mobile phase, which is a fluid mixture comprising a compound of interest for purification or separation as well as one or more solvents, is caused to flow through the column under pressure from an input end to an output end. Generally, the chemical properties of the stationary phase and the mobile phase solvents are such that the degree of partitioning of the compound of interest between the mobile phase and the stationary phase is different from the degree of partitioning of other compounds within the fluid. As a result, the degree of retention or time of retention of the compound of interest within the column is different from the degree or time of retention of the other compounds, thus causing a physical separation or at least partial purification of the compound of interest from the other compounds.

There are numerous solvents available for liquid chromatography. For instance, the HPLC solvents available under the Fluka® brand name from Sigma-Aldrich Corporation (3050 Spruce Street, St. Louis, Mo. 63103 USA) include: water, Acetonitrile, Benzene, 1-Butanol, 2-Butoxyethanol, tert-Butyl methyl ether, Carbon tetrachloride, 1-Chlorobutane, Chloroform, 2-Chloropropane, Cyclohexane, Cyclopentane, 1,2-Dichloroethane, Dichloromethane, Diethyl ether, 1,2-Dimethoxyethane, N,N-Dimethylacetamide, Dioxane, Ethanol, Ethanol, Ethyl acetate, Heptane, Hexane, Isooctane, Methanol, Methanol, Methyl acetate, Nitromethane, Pentane, 1-Propanol, 2-Propanol, 2-Propanol, Tetrachloroethylene, Tetrahydrofuran, and Toluene.

Within a chromatograph instrument or system, solvents or other reagents are generally made available to the various columns, pumps, valves and associated interconnecting tubing lines by means of a dedicated rack or compartment. The rack or compartment generally comprises a dedicated storage area for the set of solvents or other reagents that will routinely be needed or that may be needed by the chromatograph instrument or system during the course of several separations. Since many such solvents or reagents present known health hazards or are flammable, the reagent rack is generally designed to be closed during normal operation and to only be accessed by an operator or technician at such times that one or more solvents or reagents need to be replaced, having been depleted over the course of operation of the instrument or system.

Successful chromatographic separations depend on specific chemical interactions of the various analytes and other components with a stationary phase and with the various chemical constituents of a mobile phase. Because different analytes have different respective chemical properties, it is important that the correct set of solvents or reagents for an analysis at hand are mixed with a sample containing or potentially containing any particular analyte. Therefore, the various different solvents or reagents are provided in respective dedicated bottles or other containers within a reagent or solvent rack or compartment. The different containers or bottles either have respective assigned locations within the rack or compartment or are associated with different respective assigned draw tubes for aspiration of the solvent or reagent into the system.

Because of the specificity of solvents or other reagents required for any particular chromatographic analysis protocol, it is important that these materials are not confused with one another (or with completely different substances) or misplaced within a reagent or solvent rack or compartment. Although reagents, solvents and other required chemicals are generally supplied by manufacturers in well-labeled containers, these materials may be re-distributed into smaller containers within a laboratory environment. The smaller containers may be multi-purpose, initially-unlabeled vessels which require appropriate manual labeling upon initial receipt of material transferred from a manufacturer's original container. The manual label applied in a laboratory may be a non-permanent label. After manual labeling, the small transfer vessel may be handled within the laboratory many times and by many different users, since multiple replenishments from a large-volume manufacturer's “bulk” container may be required as the material within the vessel is routinely consumed. The same vessel may be re-inserted into a solvent or reagent rack many times.

Many opportunities for operator error will occur over the course of the multiple handlings of the transfer vessel or, occasionally, even a manufacturer's original container. For instance, a temporary label may be lost and replaced with an incorrect label. Even if the label is correct, the operator may transfer the wrong material into the transfer vessel. Even if the label and material are correct, the operator may mis-place the vessel within a reagent rack or compartment. Conventional chromatograph systems are designed to expect that particular solvents or reagents will be drawn into particular respective tubing lines. If an incorrect material is supplied, through any one or more of the errors listed above, the chromatograph will continue to perform the pre-programmed steps of an analysis protocol with the wrong material. This may lead to incorrect or poor-quality results, necessitating repetition of many faulty analyses. In a worst-case scenario, the error may never be discovered, and inappropriate actions may be taken—such as an incorrect treatment for a patient—based on the incorrect analytical results. Accordingly, there is a need in the art for an automated chromatograph system that can take automated procedural steps in an attempt to recognize unexpected solvents or reagents before analysis steps are performed unexpected material and that can raise an operator alert if any such errors are detected.

DISCLOSURE OF INVENTION

The present disclosure addresses the above-noted needs in the conventional art through the teaching of methods and systems for monitoring properties of fluids provided to liquid chromatography systems and comparing the monitored properties to the values that are expected if correct fluids are provided. Such methods and systems are also capable of monitoring the leak-tight worthiness of pumps and other mechanical or fluid-containing components of the liquid chromatography systems.

In accordance with a first aspect of the present teachings, there is disclosed a system for providing a solvent or reagent to a liquid chromatography system comprising: a valve comprising a common port and a plurality of other ports, configurable such that the common port may be fluidically coupled to any one of the other ports; a pump fluidically coupled to the common port of the valve; a plug configured to block flow through a first one of said other ports of the valve; a container containing the solvent or reagent, said container fluidically coupled to a second one of said other ports of the valve; a fluid tubing line having a known resistance to fluid flow fluidically coupled to a third one of said other ports of the valve; and a pressure gauge configured to measure fluid pressure within the pump, wherein the solvent or reagent is provided to the liquid chromatography system by a fourth one of the other ports. The pumps may comprise syringe pumps. The system may further comprise a computer or electronic controller electrically or electronically coupled to the pump, the pressure gauge and the valve. The system may still further comprise an electronically-readable medium having thereon program instructions readable by the computer or electronic controller, said instructions operable to cause the computer or electronic controller to measure readings of the pressure gauge while causing the pump to apply a force to a fluid therein so as to urge said fluid to either the first or third port of the valve.

In accordance with a second aspect of the present teachings, there is disclosed a method for monitoring fluids within a liquid chromatography system comprising: (a) configuring a valve so as to draw a fluid from a container into a pump; (b) configuring the valve so as to fluidically couple the pump to a port of the valve that is coupled to a plug that prevents fluid flow through said port; (c) causing the pump to progressively compress the fluid therein, while measuring a pressure of the fluid in the pump; and (d) determining if a rate of increase of the measured pressure substantially matches an expected value. Additional steps of the method may comprise: (e) upon measuring a maximum pressure, maintaining a piston of the pump in a constant position for a time of pre-determined length while continuing to measure the pressure of the fluid in the pump; and (f) determining if a decrease of the measured pressure by more than an acceptable value occurred during the time period. Still further steps of the method may comprise: (g) causing the pump to relieve the pressure of the fluid in the pump; (h) configuring the valve so as to fluidically couple the pump to a fluid pathway having a pre-determined resistance to fluid flow therethrough; (i) causing the pump to displace fluid into fluid pathway at a set flow rate while measuring the pressure of the fluid in the pump; and (j) determining if an increase of the measured pressure during the fluid displacement substantially matches a second expected value. The method may include raising an alarm either that the fluid in the container may not match expectations or that the pump may not be leak-free according to expectations depending on the measured increases or decreases in the pressure of the fluid in the pump.

In accordance with a third aspect of the present teachings, there is disclosed a liquid chromatography system comprising: (a) a mixing valve system or apparatus having an output port and a plurality of input ports thereof, each of the plurality of input ports fluidically coupled to a respective fluid-providing sub-system, each fluid-providing sub-system comprising: (i) a valve comprising a common port and a plurality of other ports, configurable such that the common port may be fluidically coupled to any one of the other ports; (ii) a pump fluidically coupled to the common port of the valve; (iii) a plug configured to block flow through a first one of said other ports of the valve; (iv) a container containing a fluid, said container fluidically coupled to a second one of said other ports of the valve; (v) a fluid tubing line having a known resistance to fluid flow fluidically coupled to a third one of said other ports of the valve; (vi) a pressure gauge configured to measure fluid pressure within the pump; and (vii) a fourth one of the other ports fluidically coupled to the mixing valve system or apparatus; (b) a chromatograph column having a first end fluidically coupled to the output of the mixing valve system or apparatus and a second end; and (c) a detector fluidically coupled to the second end of the chromatograph column.

BRIEF DESCRIPTION OF DRAWINGS

The above noted and various other aspects of the present invention will become apparent from the following description which is given by way of example only and with reference to the accompanying drawings, not drawn to scale, in which:

FIG. 1 is a schematic illustration of a generalized conventional liquid chromatography-mass (LCMS) spectrometry system;

FIG. 2 is a schematic illustration of an LCMS system in accordance with the present teachings;

FIG. 3 is an illustration of an exemplary rotary valve assembly as may be employed within an apparatus in accordance with the present teachings;

FIG. 4A is a schematic diagram of an exemplary two column LCMS system employing a fluid monitoring portion in accordance with the present teachings;

FIG. 4B is a schematic diagram of a second exemplary two column LCMS system employing a fluid monitoring portion in accordance with the present teachings;

FIG. 5 is a schematic diagram showing an example of valve configurations and fluid flow paths steps that may be employed in a chromatography method employing the system of FIG. 4A; and

FIG. 6 is a flow diagram of a method for chromatography fluid monitoring and verification in accordance with the present teachings.

MODES FOR CARRYING OUT THE INVENTION

The following description is presented to enable any person skilled in the art to make and use the invention, and is provided in the context of a particular application and its requirements. Various modifications to the described embodiments will be readily apparent to those skilled in the art and the generic principles herein may be applied to other embodiments. Thus, the present invention is not intended to be limited to the embodiments and examples shown but is to be accorded the widest possible scope in accordance with the features and principles shown and described. To appreciate the features of the present invention in greater detail, please refer to FIGS. 1-6 in conjunction with the following discussion.

FIG. 1 is a schematic illustration of a conventional liquid chromatography (LC) system. The system 10 shown in FIG. 1 comprises a chromatograph column 7 for separating a liquid chemical mixture into its constituent substances and a detector 20 (such as spectrophotometer or a mass spectrometer) fluidically coupled to the column 7 for detecting or identifying some or all of the separated constituent substances as they are received, in sequence from the column 7 The column receives a fluid stream comprising one or more selected solvent fluids or reagents supplied from containers 8 as well as a sample of interest from sample source 4. The various different solvent or reagent fluids, which may comprise a chromatographic mobile phase, are delivered along fluid tubing lines 6 a to selecting valve or mixing apparatus 9 which may mix the fluids or select a particular fluid. As illustrated, the apparatus 9 is a three-way valve but may comprise a more complex valve or valve system if more than two different solvent fluids are provided. Alternatively, the apparatus 9 could comprise a simple mixing junction or mixing chamber.

The fluids are drawn into the system 10 and propelled to the chromatographic column 7 therein by means of a pump 11 that is fluidically coupled to the output of the selecting valve or mixing apparatus 9 by fluid tubing line 6 d. Alternatively, the single illustrated fluid pump 11 could be replaced separate pumps—one for each solvent or reagent —disposed at positions 11 a in fluid tubing lines 6 a. The fluids output from the pump or pumps are delivered to a sample injector apparatus 5 along fluid tubing line 6 e and are mixed together with a sample provided from the sample source 4. The sample injector apparatus 5 may comprise, in a well-known fashion, a multiple-port rotary valve 23 and an injection loop 6 p fluidically coupled between two of the ports.

An input of the column 7 of system 10 is fluidically coupled to and receives a mixture of sample and solvent fluids from an output port of the sample injector apparatus 5 by fluid tubing line 6 f. Differential partitioning of the various chemical constituents of the mixture between the mobile phase and a stationary phase packed within the column leads to differential retention of the various constituents within the column and consequent different respective times of elution of the constituents from the column output to fluid tubing line 6 g. An optional valve 12 may separate the eluting substances, either continuously or at various times, into a portion that is delivered to waste container 14 along fluid tubing line 6 w and an analysis portion that is delivered the detector 20 along fluid tubing line 6 h.

The conventional system 10 shown in FIG. 1 is susceptible to the possible handling errors of a transfer vessel or, occasionally, even a manufacturer's original container, as described supra herein. Accordingly, FIG. 2 provides a schematic illustration of an improved LCMS system, system 50, in accordance with the present teachings. In the system 50, the solvents or reagents that are drawn from containers 8 into the fluid tubing lines 6 a are delivered to a multiple-port source valve 9 r.

As indicated in the inset 33 of FIG. 2, the source valve 9 r comprises a common port p0 as well as several dedicated ports—in the illustrated system, six dedicated ports denoted as ports p1-p6. The source valve 9 r may comprise a type of a known multiple-port rotary valve, such as the valves known as Rheodyne valves sold by IDEX Health & Science, 619 Oak Street Oak Harbor, Wash. USA. A more-detailed illustration of an exemplary source valve 9 r is shown in FIG. 3. The source valve 9 r shown in FIG. 3 comprises, in known fashion, a stator portion 66 having a plurality of fluid passages 68 therein passing through the stator portion from one end to another end and a rotor portion 62 having a groove or channel 64 on a side facing the stator portion. As shown, the stator comprises a central fluid passage (corresponding to the port p0 shown in FIG. 2) as well as six peripheral fluid passages (corresponding to the ports p1-p6 shown in FIG. 2) radially disposed about the central passage. The rotor portion 62 may rotate, as illustrated by the double-headed arrow in FIG. 3, so that any one of the peripheral passages may be fluidically connected to the central passage, within the source valve 9 r, by means of the groove or channel 64. Although a rotary valve is shown, the invention is not intended to be limited to such, as the rotary valve is but one example of a valve which may be employed.

Returning to FIG. 2, the solvents or reagents are drawn from their respective containers 8 into and through the source valve 9 r by means of a syringe pump 11 s which is coupled to the central or common port p0 as well as to a pressure sensor or gauge 16. Two different solvent or reagent fluids may be fluidically coupled to two respective ports, for example, ports p5 and p6, of the source valve 9 r by means of the fluid tubing lines 6 a. To draw either one of these fluids into the cylinder of the syringe pump, the source valve is configured such that the central port p0 is fluidically coupled to one of ports p5 and p6 while a piston of the syringe pump is withdrawn.

A computer or other electronic logic controller 32 may be included within the system 50 so as to receive information from and transmit control signals to various components of the system. The computer or other electronic logic controller 32 may be electronically coupled to the pump 11 s, the pressure gauge 16 and the source valve 9 r by means of electronic communication lines 34 a, 34 b and 34 c, respectively. The computer or other electronic logic controller 32 may also be electronically coupled to other components of the system 50, although such couplings are not explicitly illustrated in FIG. 2.

One port, for example, port pl of the source valve is blocked or plugged so that fluid cannot exit through this port and may be a default position of the source valve when the pump is not in use. If the source valve is configured to dispense solvent to port p1 and force is applied to the syringe pump piston, the pressure measured by sensor or gauge 16 is expected to rise rapidly. In the absence of a leak, the rate of pressure rise depends on fluid compressibility. The rate of pressure rise may be used to verify the correctness of a particular solvent or reagent, from among a limited number of choices. The use of the plug position p1 as a source valve “output” can also be used to verify pump seal performance and pump priming. Using a calibration fluid of known compressibility, a failure of the pressure to increase as expected or an unexpected pressure decrease can indicate an apparatus defect.

Another port, for instance, port p2 of the source valve 9 r is the output to the sample injector apparatus 5 via fluid tubing line 6 d. Another port, for instance, port p3 is used to output a small portion of a previously aspirated solvent into a waste container through a calibrated length of resistive tubing 6 c. The tubing 6 c may comprise a restricted-diameter inner bore which provides a known resistance to fluid flow. If the source valve is configured to dispense solvent to port p3 and force is applied to the syringe pump piston, the solvent will be dispensed to the waste container 14 concurrent with a rise in pressure, as measured by sensor or gauge 16, that corresponds to solvent viscosity. This measured pressure rise may be used to verify solvent identity, from among a limited number of choices. The relation between pressure rise and viscosity may be calibrated by dispensing a calibration fluid having known viscosity through port p3. Finally, another port, such as port p4, is an output to the waste container 14, using least resistance (e.g. regular) fluid tubing line 6 b, which is used for pump prime and purge operations.

With regards to the system 50 shown in FIG. 2, it is to be kept in mind that the system may be expanded by including additional instances of the sub-set of components comprising: the one or more solvent or reagent containers 8, the source valve 9 r having a plugged port, the pump 11 s, the pressure gauge 16, the known-resistance tubing 6 c, the waste container 14 and the other connecting fluid tubing lines 6 a, 6 b and 6 d. This sub-set of components may be considered to comprise a fluid-providing sub-system of the liquid chromatograph system 50. Alternatively or in addition, different respective solvents may be provided in different respective instances of said fluid-providing sub-system, especially if a large number of solvents or reagents are provided within the system.

Many liquid chromatography systems employ more than one chromatographic column during fractionation, separation or purification of an analyte. For instance a first column may comprise a sample “cleanup” column and a second column may comprise an analytical column. The cleanup column, according to some embodiments, may be a size exclusion or affinity liquid chromatography column or a High-Turbulence Liquid Chromatography column used for matrix interference removal. For instance, a test sample may applied to a first column (e.g., a clean-up column such as a Cyclone P column or the like) at the inlet port, eluted with a solvent or solvent mixture onto a second column (e.g., an analytical column such as a Hypersil Gold PFP or the like), and eluted with a solvent or solvent mixture from the second column to the outlet port. Different solvent modes may be selected for eluting the analytes. For example, liquid chromatography may be performed using a gradient mode, an isocratic mode, or a polytyptic (i.e. mixed) mode.

FIGS. 4A and 4B are schematic diagrams of two exemplary two-column LCMS systems employing a fluid monitoring portion in accordance with the present teachings. Sample injector 5 and fluid tubing line 6 f are reproduced from FIG. 2. For clarity of presentation, components upstream of the sample injector (including the solvent or reagent containers 8, the source valve 9 r, the pump 11 s, the pressure gauge 16, the known-resistance tubing 6 c, the waste container 14, the connecting fluid tubing lines 6 a, 6 b and 6 d and the electronic communication lines 34 a, 34 b and 34 c) are not shown but are considered to be present in the systems shown in FIGS. 4A-4B. In both the system 70 (FIG. 4A) and the system 75 (FIG. 4B), two chromatograph columns—a first column 7 a and a second column 7 b—are utilized. The first column 7 a may advantageously comprise a cleanup column that may be employed to separate certain classes or sub-sets of compounds from one another (e.g. large molecule versus small molecule, or polar versus non polar) with the fraction that may contain possible analyte substances retained and the other fraction discarded (or vice versa). The second column, column 7 b, is an analytical column that may be similar to the single column 7 of the system 50 (FIG. 2). The retained fraction eluted from the first, cleanup column 7 a may be separated into particular isolated compounds by the second column 7 b. The eluted constituents may be provided to detector 20 along fluid tubing line 6 h.

As an example of a two-stage chromatographic separation, a TurboFlow® column (also known as a High Turbulance Liquid Chromatography or HTLC column) may be employed as the cleanup column 7 a in a first separation step in order to isolate and possibly concentrate a subset of compounds based on their size range or molecular weight range (or some other property). TurboFlow® methods and apparatus are described in detail in U.S. Pat. Nos. 5,772,874; 5,919,368 and 6,149,816, all of which are hereby incorporated by reference in their entirety as if fully set forth herein. Briefly stated, the TurboFlow® apparatus and methods include or relate to a chromatography column or body that is formed as a substantially uniformly distributed multiplicity of rigid, solid, porous particles having substantially uniform mean cross-section dimensions or diameters of not less than about 30 μm, typically 50 μm or greater up to, but not limited to, 1000 μm in certain instances. The particles are selected from a range of various sizes and shapes and are held together in a body or column as by pressure, sintering and the like so that interstitial channels having a total interstitial volume of not less than about 45% of the total volume of the column are formed between the particles. The surfaces of the particles, including the inner surfaces of the pores in the particles, are chromatographically active, as by being coated with chromatographic stationary phase layers.

Because of the nature of the particles and packing in a TurboFlow® column, the flow of the fluid mixture through the column can be at a high flow rate and is believed that, under such conditions, turbulent flow of the mixture is induced within at least a major portion of the interstitial volume, and it is postulated that such turbulent flow in fact enhances the rate of mass transfer, thus increasing the dynamic capacity of the column. From the principles of turbulence, diffusion, and chemistry, small sample molecules may be separated from a sample matrix in a TurboFlow® column. Since small molecular weight molecules diffuse faster than large molecular weight molecules, the small sample compounds diffuse into the particle pores. The turbulent flow of the mobile phase quickly flushes the large sample compounds through the column to waste before they have an opportunity to diffuse into the particle pores. Of the sample molecules that enter the pores, those that have an affinity to the chemistry inside the pores bind to the internal surface of the column particles. The small sample molecules that have a lower binding affinity quickly diffuse out of the pores and are flushed to waste. A change in mobile phase, temperature or other parameter may then cause those molecules that were bound by the TurboFlow® column to elute to the analytical column for further separation.

The flow of analyte bearing or other fluids—including samples, solvents and mixtures thereof possibly together with other chemical components—through the two chromatograph columns 7 a, 7 b of either the system 70 or the system 75 is controlled by two multi-port valves v1, v2. Each valve may be a rotary valve of a known type, such as Rheodyne valves in which a rotor portion comprises two or three channels that may fluidically interconnect various pairs of adjacent ports, depending on the orientation of the rotor portion. The rotation and channels are schematically indicated, respectively, by a double-headed arrow and by a set of dotted straight lines in each of the valves v1 and v2. The first valve v1 may be configured to as to fluidically interconnect the members of three different pairs of adjacent ports; the second valve may be configured so as to fluidically interconnect a first pair of adjacent ports as well as all the ports of a triplet of ports, as shown by the dotted line.

One port of the first valve v1 receives a fluid from fluid tubing line 6 f. Fluid tubing lines 6 j and 6 k fluidically connect the ports of the first column 7 a to respective ports of the first valve; fluid tubing line 6 n fluidically connects a port of the first valve v1 to a port of the second valve v2 and another fluid tubing line 6 m fluidically interconnects two ports of the first valve. The first valve v1 and associated fluid tubing lines may be configured (as shown) such that fluid may be caused to flow through the first column 7 a in either direction.

In either the system 70 or the system 75, a port of the second valve receives a fluid, possibly comprising various solvents or other chemical constituents or mixtures thereof, from a solvent source 3 via fluid tubing line 6 s. The solvent source 3 may comprise various reagent containers, a rotary source valve, a syringe pump, a pressure gauge, a resistive fluid tubing line, a waste container and other interconnecting fluid tubing lines respectively similar to corresponding features illustrated in FIG. 2. Thus the solvent source 3 may include a second parallel instance of a fluid-providing sub-system as described with reference to the system 50. One or more electronic communication lines 34 f may electronically couple various of the components of the solvent source 3 to the computer or other electronic logic controller 32 in similar fashion as shown in FIG. 2.

Two ports of the second valve v2 may be plugged or otherwise unused, as indicated by hatch marks in FIGS. 4A, 4B. Another port of valve v2 directs non-analyzed fluids to a waste container 14 along fluid tubing line 6 w. A final port of the valve v2 is fluidically coupled to an inlet port of the second chromatograph column 7 b via fluid tubing line 6 q. One or more electronic communication lines 34 g may couple the computer or other electronic logic controller 32 to the valves v1, v2 so as to control their operation.

The system 75 illustrated in FIG. 4B is similar to the system 70 illustrated in FIG. 4A with the exception that the two chromatograph columns 7 a, 7 b are housed together in a cartridge or housing 40. An example of a two-column cartridge that may be employed as the cartridge 40 is disclosed in a co-pending International (PCT) Application filed on even date herewith titled “Modular Multiple-Column Chromatography Cartridge” (Attorney Docket No. 5854WO1/PCT; Application No. PCT/US2011/058229, Published as WO 2012/058515 A2) and assigned to the assignee of the present invention and incorporated herein by reference in its entirety. The cartridge 40 may include, in addition to a housing, a computer-readable identification (an indicator or identifier), such as a barcode or an RFID tag and may also include on-board computer readable memory, such as flash memory or any other form of electronic memory device, as well as an on-board electronic processor. The on-board memory, if present, may be used to store data relating to the use of the columns of the cartridge, such as supported chromatographic methods or column usage history. Further, the cartridge 40 may include one or more heaters to maintain one or the other of the columns at a temperature corresponding to an analysis protocol, as well as one or more sensors of temperature or some other physical quantity. Accordingly, the cartridge may be electronically coupled to the computer or other electronic logic controller 32 by an electronic communication line 34 h so as to, for instance, transfer identification or other data to or from a memory unit of the cartridge, to control heaters or to monitor the sensors.

FIG. 5 is a schematic diagram showing an example of valve configurations and fluid flow paths steps that may be employed in a chromatography method employing the system of FIG. 4A. The top, middle and bottom diagrams of FIG. 5 respectively illustrate: a sample loading step in which an analyte-bearing fluid is delivered to the first column 7 a; a transfer step in which the at least partially purified analyte is mixed with a solvent in valve v2 and transferred to the second column 7 b; and an eluting step, in which the analyte is eluted from the second column 7 b and transferred to the detector (not shown in FIG. 5). In these diagrams, different flow paths are distinguished from one another by lines of different appearance (i.e., solid, dotted, dashed and dash-dot lines). Other modes of operation, utilizing alternative sets of valve configurations or sequences, are also possible.

FIG. 6 (extending over FIG. 6A and FIG. 6B) is a flow diagram of a method 100 for chromatography fluid monitoring and verification in accordance with the present teachings. The method 100 may be executed by software or firmware of the computer or other electronic logic controller 32 in conjunction with signals transmitted along the electronic communication lines 34 a, 34 b and 34 c. In the first step, Step 102 of the method 100, a user selects an LC method which includes solvents or reagents for which compressibility and viscosity information is available. In the next step, Step 104, a valve, such as the source valve 9 r, is selected such that a pump (for instance, the syringe pump 11 s) draws a fluid from a designated solvent or reagent bottle. In Step 106, the valve is configured so as fluidically couple the pump cylinder, filled with the fluid drawn in step 104, to a plugged location, such as location pl of the source valve 9 r. In Step 108, the pump is operated so as to compress the fluid therein, while monitoring pump pressure, such as with gauge 16. Then, in the decision step, Step 110, if the observed rate of pressure increase (from Step 108) does not substantially match an expected value for an expected fluid, then the method execution is caused to branch to an execution termination or interruption step, Step 112, in which an alarm is raised that the solvent in the location from which the fluid was drawn may not match expectations. Upon observing the alarm, a user may perform any appropriate tests or checks to determine if the correct solvent or reagent is loaded in the position from which fluid was drawn. Depending upon the results of such tests or checks, the user may replace the solvent or reagent and re-start execution of the method 100 or, alternatively, may re-set the alarm (possibly, after solvent or reagent replacement) and continue execution of the method from the interrupted point.

If, during execution of step 108 of the method 100 (FIG. 6), the observed rate of pressure increase does indeed substantially match an expected value for an expected fluid, then step 110 branches execution to Step 114 in which, after obtaining a maximum useable pressure, the pump piston is held in place for a set amount of time while continuing to measuring any change or changes in pressure. In the subsequent decision step, Step 116, if an observed pressure drop is greater than an than an acceptable amount during the selected time, then execution is caused to branch to an execution termination or interruption step, Step 118, in which an alarm is raised that the pump in the given location may not be leak-free in accordance with expectations. Upon observing the alarm, a user may perform any appropriate tests or checks to determine if the pump is operating correctly. Depending upon the results of such tests or checks, the user may need to replace or repair the pump or other components and re-start execution of the method 100 or, alternatively, may re-set the alarm and continue execution of the method from the interrupted point.

The method 100 (FIG. 6) branches to Step 120 if, in Step 116, the observed pressure drop is determined to not exceed the acceptable amount during the selected time. In step 120, the pump may be operated, such as by moving a pump piston, so as to relieve the pressure in pump chamber. In the subsequent Step 122, the valve, such as source valve 9 r, is operated so as to fluidically connect the pump with a fluid pathway having a known fluid resistance; in Step 124, the pump is operated so as to inject the fluid into such pathway at a set flow rate while the pump pressure is measured. In the subsequent decision step, Step 126, if the observed pressure increase (of Step 124) does not match an expected value, then execution is caused to branch to a termination or interruption step, Step 128, in which an alarm is raised that that the solvent in the location from which the fluid was drawn may not match expectations. Upon observing the alarm, a user may perform any appropriate tests or checks to determine if the correct solvent or reagent is loaded in the position from which fluid was drawn. Depending upon the results of such tests or checks, the user may replace the solvent or reagent and re-start execution of the method 100 or, alternatively, may re-set the alarm (possibly, after solvent or reagent replacement) and continue execution of the method from the interrupted point.

Execution of the method 100 proceeds to Step 130 if all pressure monitoring tests have yielded acceptable results. At this point, it may be reported to a user that the pump and solvent check passed with acceptable measurements. Subsequently, the valve may be configured so as to dispense the solvent or reagent into the system along fluid tubing line 6 f (if the solvent is to be utilized) or to waste.

An improved liquid chromatography system has been disclosed. Advantageously, a system in accordance with the present teachings may be employed in an automated sample preparation and analysis system, such as is disclosed in a co-pending U.S. Provisional application for patent titled “Automated System for Sample Preparation and Analysis” (Attorney Docket No. TFS-13, Application No. 61/408,180) filed on Oct. 29, 2010. In various embodiments, the automated sample preparation and analysis system includes a sample preparation system for preparing various samples and a sample analysis system, which may include a liquid chromatography mass spectrometer (“LCMS”) for analyzing the prepared samples according to selected analyte assays. The sample preparation system and the sample analysis system are interconnected in an automated manner. The automated sample preparation and analysis system is designed to generally operate with minimal operator intervention or maintenance and includes at least one controller for, inter alia, controlling valve configurations and, optionally, monitoring operational or instrumental conditions. Because of the automated nature of the instrument, it is advantageous for the automated system to be able to monitor its own configuration and operating state and to provide an alert an operator if the system detects any possible problems. A system for liquid chromatography in accordance with the present teachings may assist in these functions.

The discussion included in this application is intended to serve as a basic description. Although the present invention has been described in accordance with the various embodiments shown and described, one of ordinary skill in the art will readily recognize that there could be variations to the embodiments and those variations would be within the spirit and scope of the present invention. The reader should be aware that the specific discussion may not explicitly describe all embodiments possible; many alternatives are implicit. Accordingly, many modifications may be made by one of ordinary skill in the art without departing from the spirit, scope and essence of the invention. Neither the description nor the terminology is intended to limit the scope of the invention. All patent application publications or other publications are hereby explicitly incorporated by reference herein as if fully set forth herein. 

What is claimed is:
 1. A system for providing a solvent or reagent to a liquid chromatography system comprising: a valve comprising a common port and a plurality of other ports, configurable such that the common port may be fluidically coupled to any one of the other ports; a pump fluidically coupled to the common port of the valve; a plug configured to block flow through a first one of said other ports of the valve; a container containing the solvent or reagent, said container fluidically coupled to a second one of said other ports of the valve; a fluid tubing line having a known resistance to fluid flow fluidically coupled to a third one of said other ports of the valve; and a pressure gauge configured to measure fluid pressure within the pump, wherein the solvent or reagent is provided to the liquid chromatography system by a fourth one of the other ports.
 2. A system as recited in claim 1, further comprising a second container containing a second solvent or reagent fluidically coupled to a fifth one of said other ports of the valve, said system configurable so as to also provide the second solvent or reagent to the liquid chromatography system.
 3. A system as recited in claim 1, wherein the pump comprises a syringe pump.
 4. A system as recited in claim 1, further comprising a computer or electronic controller electrically or electronically coupled to at least one of the pump, the pressure gauge and the valve.
 5. A system as recited in claim 1, further comprising a computer or electronic controller electrically or electronically coupled to the pump, the pressure gauge and the valve.
 6. A system as recited in claim 1, further comprising an electronically-readable medium having thereon program instructions readable by the computer or electronic controller, said instructions operable to cause the computer or electronic controller to measure readings of the pressure gauge while causing the pump to apply a force to a fluid therein so as to urge said fluid to either the first or third port of the valve.
 7. A system as recited in claim 6, wherein the program instructions are further operable to cause the computer or electronic controller to compare increases or decreases of the measured readings of the pressure gauge to expected values.
 8. A method for monitoring fluids within a liquid chromatography system comprising: (a) configuring a valve so as to draw a fluid from a container into a pump; (b) configuring the valve so as to fluidically couple the pump to a port of the valve that is coupled to a plug that prevents fluid flow through said port; (c) causing the pump to progressively compress the fluid therein, while measuring a pressure of the fluid in the pump; and (d) determining if a rate of increase of the measured pressure substantially matches an expected value.
 9. A method for monitoring fluids within a liquid chromatography system as recited in claim 8, further comprising: (e) raising an alarm that the fluid in the container may not match expectations if the rate of increase of the measured pressure does not substantially match the expected value.
 10. A method for monitoring fluids within a liquid chromatography system as recited in claim 8, further comprising: (e) upon measuring a maximum pressure, maintaining a piston of the pump in a constant position for a time of pre-determined length while continuing to measure the pressure of the fluid in the pump; and (f) determining if a decrease of the measured pressure by more than an acceptable value occurred during the time period.
 11. A method for monitoring fluids within a liquid chromatography system as recited in claim 10, further comprising: (g) raising an alarm that the pump may not be leak-free according to expectations if the decrease of the measured pressure during the time period is greater than the acceptable value.
 12. A method for monitoring fluids within a liquid chromatography system as recited in claim 10, further comprising: (g) causing the pump to relieve the pressure of the fluid in the pump; (h) configuring the valve so as to fluidically couple the pump to a fluid pathway having a pre-determined resistance to fluid flow therethrough; (i) causing the pump to displace fluid into fluid pathway at a set flow rate while measuring the pressure of the fluid in the pump; and (j) determining if an increase of the measured pressure during the fluid displacement substantially matches a second expected value.
 13. A method for monitoring fluids within a liquid chromatography system as recited in claim 12, further comprising: (k) either raising an alarm that the fluid in the container may not match expectations if the rate of increase of the measured pressure does not substantially match the second expected value or reporting that the pump performance and fluid properties substantially match expectations.
 14. A liquid chromatography system comprising: (a) a mixing valve system or apparatus having an output port and a plurality of input ports thereof, each of the plurality of input ports fluidically coupled to a respective fluid-providing sub-system, each fluid-providing sub-system comprising: (i) a valve comprising a common port and a plurality of other ports, configurable such that the common port may be fluidically coupled to any one of the other ports; (ii) a pump fluidically coupled to the common port of the valve; (iii) a plug configured to block flow through a first one of said other ports of the valve; (iv) a container containing a fluid, said container fluidically coupled to a second one of said other ports of the valve; (v) a fluid tubing line having a known resistance to fluid flow fluidically coupled to a third one of said other ports of the valve; (vi) a pressure gauge configured to measure fluid pressure within the pump; and (vii) a fourth one of the other ports fluidically coupled to the mixing valve system or apparatus; (b) a chromatograph column having a first end fluidically coupled to the output of the mixing valve system or apparatus and a second end; and (c) a detector fluidically coupled to the second end of the chromatograph column.
 15. A liquid chromatography system as recited in claim 14, wherein each of the pumps comprises a syringe pump.
 16. A liquid chromatography system as recited in claim 14, further comprising: (d) a computer or electronic controller electrically or electronically coupled to the pump, the pressure gauge and the valve of at least one fluid-providing sub-system.
 17. A liquid chromatography system as recited in claim 16, further comprising: (e) an electronically-readable medium having thereon program instructions readable by the computer or electronic controller, said instructions operable to cause the computer or electronic controller to measure readings of the pressure gauge of a one of the fluid-providing sub-systems while causing the pump of the one of the fluid-providing sub-systems to apply a force to a fluid therein so as to urge said fluid to either the first or third port of the valve of the one of the fluid-providing sub-systems. 